Miyamoto Ei, Ohsumi Akihiro, Hwang David, Boonstra Kristen, Cypel Marcelo, Martinu Tereza, Keshavjee Shaf, Juvet Stephen C
Latner Thoracic Research Laboratories, University Health Network, Toronto, Ontario, Canada.
Department of Pathology, Sunnybrook Hospital, Toronto, Ontario, Canada.
JHLT Open. 2025 May 26;9:100299. doi: 10.1016/j.jhlto.2025.100299. eCollection 2025 Aug.
Ex vivo lung perfusion (EVLP) has been clinically applied as a lung preservation and assessment tool prior to lung transplantation (LTx) and is evolving to become a platform to deliver cellular and gene therapies or inactivate pathogens. Here we aimed to investigate the utility of our recently reported rat EVLP-to-LTx model as the smallest ever experimental survival model of EVLP-to-LTx and to compare late graft endpoints between strain combinations.
We tested three strains as normothermic EVLP donors: Fisher 344 (F344), Lewis (LEW), and Wistar Kyoto (WKy) rats. Then we tested three strain combinations of EVLP-to-LTx (F344-to-WKy, F344-to-LEW, and LEW-to-LEW) to compare histologic and radiologic changes.
F344 and LEW, but not WKy rat lungs, tolerated 4 hours of normothermic EVLP. F344-to-WKy EVLP-to-LTx developed significant histologic (as measured by acute lung injury score, ISHLT A and B grade rejection score) and radiologic (volume and mean Hounsfield units of aerated lung graft analyzed by computed tomography at day 7 after EVLP-to-LTx) changes in the lung allograft. In this strain combination, progressive deterioration with time was noted up to day 28, while F344-to-LEW grafts exhibited only mild injuries similar to LEW-to-LEW. In addition, flow cytometric analyses of F344-to-WKy EVLP-to-LTx revealed a sharp rise in activation marker expression in lung graft T cells beginning at day 3.
Our F344-to-WKy EVLP-to-LTx model generates reproducible and clinically relevant histological, radiological, and immunological results similar to those seen in humans. The model is therefore well suited to experimental EVLP studies with long-term follow-up prior to moving to large animal and human studies.
体外肺灌注(EVLP)已在临床上用作肺移植(LTx)前的肺保存和评估工具,并且正在发展成为一个提供细胞和基因治疗或灭活病原体的平台。在此,我们旨在研究我们最近报道的大鼠EVLP至LTx模型作为有史以来最小的EVLP至LTx实验性存活模型的效用,并比较不同品系组合之间的晚期移植终点。
我们测试了三种品系作为常温EVLP供体:Fisher 344(F344)、Lewis(LEW)和Wistar Kyoto(WKy)大鼠。然后我们测试了三种EVLP至LTx的品系组合(F344至WKy、F344至LEW和LEW至LEW),以比较组织学和放射学变化。
F344和LEW大鼠的肺,但WKy大鼠的肺不能耐受4小时的常温EVLP。F344至WKy的EVLP至LTx在肺移植中出现了显著的组织学(通过急性肺损伤评分、国际心脏和肺移植学会A和B级排斥评分测量)和放射学(在EVLP至LTx后第7天通过计算机断层扫描分析充气肺移植的体积和平均亨氏单位)变化。在这种品系组合中,直到第28天观察到随时间的渐进性恶化,而F344至LEW移植仅表现出与LEW至LEW相似的轻度损伤。此外,对F344至WKy的EVLP至LTx进行的流式细胞术分析显示,从第3天开始,肺移植T细胞中激活标志物表达急剧上升。
我们的F344至WKy的EVLP至LTx模型产生了与人类相似的可重复且与临床相关的组织学、放射学和免疫学结果。因此,该模型非常适合在进行大型动物和人体研究之前进行长期随访的实验性EVLP研究。